KR19980076912A - 3-heterocyclic mercapto-4-hydroxyquinolin-2 (1H) -one derivatives acting as NMDA receptor antagonists and methods for their preparation - Google Patents

Abstract

The present invention is directed to a 3-heterocyclic mercapto-4-hydroxyquinolin-2 (1H) -one derivative of formula (1), a method for preparing the same, and a strychinine-insensitive glycine binding site in an NMDA receptor complex. It relates to the use as a potent and specific antagonist in action. The compounds of the present invention are useful for the treatment and prevention of neurodegenerative diseases. In particular, the compounds of the present invention are useful for reducing damage to the central nervous system resulting from anemia or hypoxia such as stroke, hypoglycemia, ischemia, cardiac arrest, trauma. In addition, the compounds of the present invention are useful for the prevention and treatment of chronic neurodegenerative diseases including epilepsy, Alzheimer's disease, Huntington's disease and Parkinson's disease. In addition, the compounds of the present invention are used as anticonvulsants, analgesics, antidepressants, anti-anxiety agents and antischizophrenia.

[Formula 1]

(R ₁ , R ₂ , R ₃ , R ₄ and R are as described in the specification.)

Description

3-heterocyclic mercapto-4-hydroxyquinolin-2 (1H) -one derivatives acting as NMDA receptor antagonists and methods for their preparation

The present invention relates to a 3-heterocyclic mercapto-4-hydroxyquinolin-2 (1H) -one derivative of formula (I), which acts as an antagonist of excitatory amino acids, a preparation thereof, and a use as a therapeutic agent for neurological diseases.

In particular, the compounds of the present invention antagonize the excitatory activity of NMDA receptors and are particularly useful for reducing damage to the central nervous system resulting from anemia or hypoxia, such as stroke, hypoglycemia, ischemia, cardiac arrest, trauma.

In addition, the compounds of the present invention are useful for preventing chronic neurodegenerative diseases including epilepsy, Alzheimer's disease, Huntington's disease and Parkinson's disease. The compounds of the present invention are also used as anticonvulsants, analgesics, antidepressants, anti-anxiety agents and antischizophrenia.

In recent years, as the elderly population increases, chronic neurodegenerative diseases such as senile dementia have increased significantly, which is becoming a social problem. Currently, about 4 million Americans aged 65 or older have senile dementia, a type of chronic neurodegenerative disease, and two-thirds of them are unable to maintain normal social life. Geriatric dementia is a gradual decrease in memory and thinking ability. About 55% of senile dementia is Alzheimer's disease. The symptoms of Alzheimer's disease are the loss of memory and the forgetting of communication methods. Recent studies on Alzheimer's disease have suggested various causes, and one of the most important causes of neuronal cell death is the abnormal activity of neurotransmitters. It has been found to be due to abnormalities occurring during differentiation, growth and degeneration processes.

L-glutamate is the most important excitatory neurotransmitter in mammalian central nervous system neurotransmission. Nearly all central nervous system neurons are excited by L-glutamate, which acts on several receptor-ion channel complexes on the surface of neurons to open ion channels and induce cations such as sodium or calcium. To excite nerve cells by inducing polarization between nerve cell membranes.

These CNS neurons have NMDA receptors excited by glutamate analogue N-methyl-D-aspartate (abbreviated as NMDA) and non-NMDA receptors that are not excited by NMDA. NMDA receptors are cell surface protein complexes widely distributed in the brain or spinal cord, and are known to be closely involved in the growth of neurons by delivering the excitability of neurons at the synapses of neurons.

One of the characteristics of this NMDA receptor is that its activity is completely blocked by Mg ⁺⁺ when the cells are at rest. However, this blockade is voltage-dependent and the block is released when the cell is activated by non-NMDA receptors or when other excitatory substances are partially nonpolarized. As such, the mechanism of action of NMDA receptors depends on conditions and plays an important role in learning and memory processes.

Another property of NMDA receptors is that the cells die when the receptor is overly excited. This may be due to excessive accumulation of Ca ⁺⁺ in cells. Recently, many studies have been conducted on cell death by excitatory toxicity of NMDA receptors.

After culturing neurons in vitro, glutamate treatment causes cells to swell and cytotoxicity. This excitatory toxicity is dependent on the presence of Ca ^{++. When} a large amount of Ca ⁺⁺ enters neurons through the ion channel of glutamate receptors, normal cell activity is destroyed and the release of glutamate is accelerated by feedback. Excitatory toxicity occurs. In this process, proteases and lipases are activated to destroy the membranes of neurons and produce free radicals that kill the cells [JW Mcdold, MV Johnson, Brain Res. Reviews 15, 41 (1990)]. As such, excitatory toxicity caused by neurotransmitters kills neurons, and research has been found to be an important cause of neurodegenerative diseases and stroke associated with brain cell death.

Recently, research has focused on the mechanism by which L-glutamate is excessively secreted by brain injury, spinal cord injury, stroke, ischemia, or hypoglycemia resulting in permanent damage to the central nervous system. It has been found that the damage caused by is prevented by NMDA receptor antagonists.

NMDA receptor antagonists prevent many clinical diseases, including ischemic symptoms and epilepsy, and prevent chronic neurodegenerative diseases such as Alzheimer's disease, Huntington's disease and Parkinson's disease [G. Johnson, Annu. Rep. Med. Chem. 24, 41 (1989); G. Johnson and C. F. Bigge, ibid. 26, 11 (1991); Werling et al., J. Pharmacol. Exp. Ther. 255, 40 (1990). Accordingly, several NMDA receptor antagonists have recently been used to treat acute stroke or brain damage.

There are also reports that NMDA receptor antagonists are useful for improving memory and learning, such as 1-aminocyclopropanecarboxylic acid methyl esters, D-cycloserine and R-(+)-3-, which are specific glycine site ligands. Amino-1-hydroxypyrrolidin-2-one- (HA-966) [Bliss, TV P, Collingride, GL, Nature 345, 347 (1990); GB 2231048A (1990).

Recent reports suggest that NMDA receptor antagonists have analgesic, antidepressant, antischizophrenic and anti-anxiety effects [Dickenson, A. H. and Aydar, E., Neuroscience Lett. 121, 263 (1990); R. Trullas and P. Skolnick, Eur. J. Pharmacol. 185, 1 (1990); J. H. Kehne, et al., Eur. J. Pharmacol. 193, 283 (1991); P. H. Hutson, et al., Br. J. Pharmacol. 103, 2037 (1991).

Non-competitive NMDA receptor antagonists such as ketamine and dextrometopan are useful for the treatment of growth disorders, autism and the like in children. NMDA receptor antagonists are also useful as protective agents in laser neurosurgery.

The fact that NMDA receptors play an important role in the synaptic degeneration of neurons, which is thought to be one of the causes of these diseases, is more evident by recent studies on their physiological function and subunit structure. Kumar KN, et al., Nature 354, 70-73 (1991); Nakanishi, S., et al., Nature 354, 31-37 (1991); Monyer, H., et al., Science 256, 1217-1221 (1992). The NMDA receptor has at least five distinct substrate binding sites, including (a) a binding site for binding to the neurotransmitter L-glutamate, (b) an allosteric modulator site for binding to glycine, and (c) in the channel Sites that bind fencyclidine and related compounds, (d) binding sites that bind Mg ⁺⁺ , and (e) binding sites that bind to the divalent cation Zn ⁺⁺ and exhibit inhibitory activity [Lynch, DR, et al., Mol. pharmacol. 45, 540-545 (1994); Kuryatov, A., et al., Neuron 12, 1291-1300 (1994); Nakanishi, S., Science 256, 1217-1221 (1992).

Looking at the physiological action of subunits of NMDA receptors, the NMDA receptors are activated by glutamate and glycine or their agonists. In addition, the associated Ca ⁺⁺ permeable channel is physiologically blocked by Mg ⁺⁺ in a voltage-dependent manner, and Zn ⁺⁺ with its own regulatory site reduces the activity of synapses of NMDA receptors [Lynch, DR, et al. , Mol. pharmacol. 45, 540-545 (1994); Kuryatov, A., et al., Neuron 12, 1291-1300 (1994); Nakanishi, S., Science 256, 1217-1221 (1992).

Pharmacological interest has focused on the NMDA receptors over the past two decades, and agonists and antagonists that depend on each of the binding sites described above have been explored as candidates for useful drugs. Among them, the most promising method for regulating NMDA receptor activity was the development of an allosteric modulator of glycine binding site.

In 1987, Johnson and Ascher discovered glycine binding sites in NMDA receptors. Subsequent studies have revealed that glycine binding sites and glutamate binding sites in NMDA receptors are present in the same protein and glycine binding sites in NMDA receptors. It was found that there is a negative allosteric coupling between and glutamate binding sites.

Because of the presence of negative allosteric coupling, binding of agonists at glutamate recognition sites reduces glycine affinity for glycine recognition sites, and antagonist binding at glutamate recognition sites is enhanced by glycine site antagonists and vice versa [ Beneveniste, M., et al. J. Physiol. 428, 333 (1990); Leser, R. A .; Tong, G. and Jahr, C. E., J. Neurosci. 13, 1088 (1993); Clements, J. D .; Westbrook, G. L., Neuron. 7, 605 (1991).

In addition, recent in vivo microdialysis (dialysis) studies have found that large amounts of glutamate were released in the ischemic brain region but little release of glycine in the rat focal ischemic model [Globus, MY T, et. al., J. Neurochern. 57, 470-478 (1991).

As a result of the experiment, it can be seen that the glycine antagonist is a very powerful neuroprotective drug that can reduce excessive excitability of neurons when neurons, neurons, are excessively excited by glutamate to cause cytotoxicity.

Glycine antagonists act non-competitively with glutamate to prevent the NMDA channels from opening, thus overcoming the high concentrations of endogenous glutamate released in the ischemic brain region, unlike competitive NMDA antagonists that compete competitively with glutamate. There is no need to do it. Thus, NMDA receptors are regulated rather than completely inhibited by glycine site antagonists, and these regulatory actions may be much more physiological than receptor blockade where receptor function is completely inhibited (compared to channel blockers), thus glycine antagonists Has fewer side effects than other antagonists. In fact, glycine antagonists can be injected directly into the brains of rodents without side effects [Tricklebank, M. D., et al., Eur. J. Pharmacol. 167, 127 (1989); Koek, W., et al., J. Pharmacol. Exp. Ther. 245, 969 (1989); Willets and Balster, Neuropharmacology 27, 1249 (1988).

As such, glycine site antagonists are noncompetitive antagonists that regulate rather than block NMDA receptors, and thus have fewer side effects from NMDA receptor blockade than other forms of NMDA receptor antagonists. It has emerged as a target substance.

The biggest problem in developing glycine-ligand compounds that react with NMDA associated glycine sites was that most of these compounds did not cross the Blood-Brain Barrier. As a result of our efforts to develop glycine-site ligands that can cross the blood-brain-membrane, we have shown that kynureric acid derivatives (2-carboxyquinoline), 2-carboxyindole derivatives, quinoxaline derivatives and 2- Quinolone derivatives and the like have been developed.

Recently, 4-hydroxyquinolin-2 (1H) -one derivatives with strong in vivo activity as selective noncompetitive antagonists of NMDA receptors have been reported. They are also active upon oral administration as glycine receptor antagonists [McOuaid, L. A., et al., J. Med. Chem. 35, 3423 (1992); Leeson, P. D., et al., J. Med. Chem. 36, 3386 (1993); Kulagowski, J. J., et al., J. Med. Chem. 37, 1402 (1994); Cai, S. X., et al., J. Med. Chem. 39, 4682 (1996); and 39, 3248 (1996); EP 489,458; EP 459,561; EP 685,466 A1; WO 94/20470; WO 93/10783, EP 685,466 A1 and EP 481,676 A1. It has been reported to be a useful drug for preventing or treating neurodegenerative diseases, convulsions and schizophrenia in general.

Glycine site antagonists have emerged as central nervous system agonist candidates because they have a wide therapeutic window between the desirable effects of neuroprotection and the side effects of hyperactivity commonly found in competitive glutamate antagonists or channel blockers.

If you look at the antagonists developed so far, there is 2-carboxyindole acrylamide developed by Glaxo,

Quinoxaline-2,3-dione, developed by Ciba-Geigy,

Shiva further refined this and developed the following compounds.

And

There is also the following compound developed by Zeneca

There is the following compound developed by Pfizer,

The following compounds were developed by Ciba-Geigy.

And

In addition, various substances have been developed, but until now, as a glycine site NMDA receptor antagonist having a large receptor affinity, it is well absorbed into the central nervous system and high solubility materials have not been developed.

The present inventors have intensively studied pharmacologically useful aspects of 2-quinolone derivatives, which have strong and specific antagonists that act on the strikinin-insensitive glycine binding site in the NMDA receptor complex, resulting in 3-hetero. In the cyclic mercapto-4-hydroxyquinolin-2 (1H) -one derivative, a compound showing strong antagonism with respect to the NMDA receptor, penetrating well into the central nervous system and having high solubility was synthesized.

It is an object of the present invention to provide a 3-heterocyclic mercapto-4-hydroxyquinolin-2 (1H) -one derivative which acts as an antagonist of excitatory amino acids, wherein the compound of the present invention is used for treating or preventing neurodegenerative diseases. Useful for

The present invention relates to 3-heterocyclic mercapto-4-hydroxyquinolin-2 (1H) -one derivatives of the general formula (1), and includes tautomeric isomers or pharmaceutically acceptable salts thereof.

Formula 1

In the above, R represents a functional group of the formula (2) or (3).

[Formula 2]

[Formula 3]

In Formula 2 or Formula 3 above, the closed circle represents two non-adjacent double bonds at any position of the pentagonal ring. A, B, C, and D independently represent oxygen, sulfur, nitrogen, or carbon, at least one of A, B, C, and D represents oxygen or sulfur, and at least one of A, B, C, and D represents carbon And others. E, F, G, H and I independently represent oxygen, sulfur, nitrogen or carbon, at least one of E, F, G, H and I represents oxygen or sulfur and E, F, G, H and I At least one of them represents something other than carbon.

R ₁ , R ₂ , R ₃ and R ₄ are independently hydrogen, hydroxy, halogen, nitro, amino, haloalkyl, cyano, alkyl, alkenyl, alkynyl, azido, acylamino, aryl, alkoxy or hetero Represents a cycle ring.

R ₅ and R ₆ are independently hydrogen, halogen, nitro, amino, carboxy, thiol, haloalkyl, cyano, alkyl, alkenyl, alkynyl, azido, acylamino, sulfonyl, aminosulfonyl, aryl, alkoxy , Heterocycle ring, acyloxy, alkylthio, arylthio, alkyl ester, alkyl carboxylate, aryl ester, aryl carboxylate, aralkyl ester, aralkyl carboxylate, urea or amidine.

Preferred compounds in the form of formula (1) are those in which R ₁ is hydrogen, nitro, amino, chloro, bromo or alkyl; R ₂ is hydrogen, chloro, bromo or trifluoromethyl; R ₃ is nitro, chloro, bromo, trifluoromethyl or alkyl; R ₄ is hydrogen, nitro, chloro, bromo or amino.

Typical aryl (C ₆ -C ₁₄ ) groups include phenyl, naphthyl, phenanthryl, anthracyl, indenyl, azulenyl, biphenyl, biphenylenyl and fluorenyl groups.

Typical amino groups include NH ₂ , NHR ₇ and NHR ₈ , wherein R ₇ and R ₈ are C ₁ -C ₄ alkyl groups.

Typical halogen atoms include fluorine, chlorine, bromine and iodine.

Typical alkyl (C ₁ -C ₄ ) groups include methyl, ethyl, propyl, isopropyl, butyl, secondary-butyl and tert-butyl groups.

Typical alkenyl (C ₂ -C ₄ ) groups include vinyl, allyl, 1-butenyl, 2-butenyl, 3-butenyl and isobutenyl groups.

Typical alkynyl (C ₂ -C ₄ ) groups include propazyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, and 3-butynyl groups.

Typical haloalkyl groups are C ₁ -C ₄ alkyl groups substituted with one or more fluorine, chlorine, bromine or iodine atoms, for example fluoromethyl, difluoromethyl, trifluoromethyl, pentafluoroethyl, 1,1-di Fluoroethyl and trichloromethyl groups.

Typical alkoxy groups include oxygen substituted by one C ₁ -C ₄ alkyl group mentioned above.

Typical heterocyclic groups include C ₃ -C ₇ heterocycloalkyl, C ₃ -C ₇ heterocycloalkyl (C ₁ -C ₆ ) alkyl, heteroaryl and heteroaryl (C ₁ -C ₆ ) alkyl; Suitable heterocycloalkyl groups include piperidyl, piperazinyl and morpholinyl groups; Suitable heteroaryl groups are pyridyl, quinolyl, isoquinolyl, pyridazinyl, pyrimidinyl, pyrazinyl, pyrrolyl, indolyl, pyranyl, furyl, benzofuryl, thienyl, benzthienyl, imidazolyl, Oxdiazolyl, thiazolyl and thiadiazolyl groups.

For use in medicine, the salts of the compounds of formula 1 should be non-toxic and pharmaceutically acceptable salts. Various salts may be used to prepare the compounds of the invention or their toxic and pharmaceutically acceptable salts.

Pharmaceutically acceptable salts of compounds of formula 1 include alkali metal salts such as lithium, sodium, or potassium salts, alkaline earth metal salts such as calcium or magnesium salts, and suitable organic ligands. Salts formed, for example, quaternary ammonium salts. Acid addition salts can be prepared by mixing a solution of a compound of the invention with a solution of a pharmaceutically acceptable non-toxic acid such as hydrochloric acid, fumaric acid, maleic acid, succinic acid, acetic acid, citric acid, tartaric acid, carbonic acid or phosphoric acid. have.

The present invention includes prodrugs of compounds of formula 1 within the scope of the present invention. In general, such prodrugs are functional derivatives of the compound of formula (I), and should be able to be readily changed to the compound required to enter the body and exhibit efficacy. Conventional procedures for the selection and preparation of suitable prodrug derivatives are described in the existing literature [Design of Prodrug, ed. H. Bundgaard, 1985].

If the compound corresponding to the present invention has at least one asymmetric center, an enantiomer may be present, and if the compound corresponding to the present invention has two or more asymmetric centers, the diastereoisomer May be present. Such isomers and mixtures thereof are included within the scope of the present invention.

Particularly preferred compounds in the present invention include the following compounds. However, the following compounds are illustrative of the present invention and the present invention is not limited to the following compounds.

3- (benzothiazol-2-ylsulfanyl) -7-chloro-4-hydroxy-1H-quinolin-2-one,

3- (benzoxazol-2-ylsulfanyl) -7-chloro-4-hydroxy-1H-quinolin-2-one,

7-chloro-4-hydroxy-3- (1-methyl-1H-imidazol-2-ylsulfanyl) -1H-quinolin-2-one,

3- (1H-benzoimidazol-2-ylsulfanyl) -7-chloro-4-hydroxy-1H-quinolin-2-one,

7-chloro-4-hydroxy-3- (1H- [1.2.4] triazol-3-ylsulfanyl) -1H-quinolin-2-one,

5,7-dichloro-4-hydroxy-3- (nicotin-2-ylsulfanyl) -1H-quinolin-2-one,

5,7-dichloro-4-hydroxy-3- (nicotin-6-ylsulfanyl) -1H-quinolin-2-one,

5,7-dichloro-4-hydroxy-3- (5-N-benzylcarbamoyl-pyridin-2-ylsulfanyl) -1H-quinolin-2-one,

7-chloro-4-hydroxy-3- (nicotin-2-ylsulfanyl) -1H-quinolin-2-one,

5,7-dichloro-4-hydroxy-3- (1H- [1.2.4] triazol-3-ylsulfanyl) -1H-quinolin-2-one and salts and prodrugs thereof.

The pharmaceutical compositions of the invention are preferably in unit dosage form such as tablets, capsules, powders, granules, sterile solutions or suspensions, or suppositories for oral, intravenous, parenteral or rectal administration. To prepare solid compositions such as tablets, the main active ingredient is a pharmaceutical carrier, e.g., the conventional tableting ingredients cornstarch, lactose, sucrose, sorbitol, talc, stearic acid, magnesium stearate, di A solid preformulation composition comprising a homogeneous mixture of a compound of the invention or a pharmaceutically acceptable non-toxic salt thereof, in admixture with calcium phosphate or gums and other pharmaceutical diluents such as water Form. The homogeneous preliminary solid composition allows the active ingredient to be dispersed evenly throughout the composition so that the composition can be easily divided into effective unit dosage forms containing the same amount such as tablets, pills and capsules. This preliminary solid composition is further subdivided into unit dosage forms of the abovementioned type comprising from 0.1 to 500 mg of the active ingredient of the invention. Tablets or pills of the novel compositions may be coated or synthesized to provide an advantageous dosage form when sustained release is required. For example, a tablet or pill may be composed of an internal dosage ingredient and an external dosage ingredient, in which the external dosage ingredient surrounds the internal dosage ingredient. The two components can be separated by an enteric membrane, which prevents the digestion of the stomach and allows the internal components to pass through the duodenum or delay the release of the internal components intact. A variety of materials are used as such enteric membranes or coating materials, which include a plurality of polymeric acids and mixtures of polymeric acids and materials such as shellac, cetyl alcohol, cellulose acetate. The liquid form of the novel compositions of the present invention prepared for oral or injectable administration consists of an aqueous solution, a properly flavored syrup, an aqueous or oily suspension and an edible oil such as cottonseed oil, sesame oil, coconut oil or 45 peanut oil. Emulsions, elixirs and similar pharmaceutical vehicles. Suitable dispersing or suspending agents for making aqueous suspensions include synthetic or natural gums such as tragacanth, acacia, alginate, dextran, sodium carboxymethylcellulose, methylcellulose, polyvinylpyrrolidone or gelatin. ). In the treatment of neurodegeneration, a suitable dosage is 0.01-250 mg / kg per day, preferably 0.05-100 mg / kg per day, more preferably 0.05-5 mg / kg per day. The compound may be conveniently administered by intravenous injection.

Hereinafter will be described a method for preparing a compound of the present invention.

The compound of Chemical Formula 1, which is a compound of the present invention, may be prepared by cyclization of a compound of Chemical Formula 4.

[Formula 4]

Wherein R ₁ , R ₂ , R ₃ , R ₄ and R are as mentioned above and A ₁ represents a reactive carboxylate functional group. The reaction is carried out in the presence of a base followed by mild acidic treatment [J. Heterocycl. Chem., 1975, 12, 351]. Suitable bases used in the reaction include Na metal, K metal, sodium hydride and potassium hexamethyldisilazide.

Suitable reactive carboxylate functional groups A ₁ are esters such as C ₁ to C ₄ alkyl esters; Acid anhydrides such as anhydrides mixed with C ₁ to C ₄ alkanoic acid; Acid halides such as acid chlorides; Orthoesters; And primary, secondary and tertiary amides. Preferably, A ₁ represents methoxycarbonyl or ethoxycarbonyl.

The intermediate of Chemical Formula 4 is prepared by reacting the compounds of Chemical Formulas 5 and 6.

[Formula 5]

[Formula 6]

In the above, R ₁ , R ₂ , R ₃ , R ₄ , R and A ₁ are as mentioned above, and X represents a halogen atom such as chlorine or bromine. The reaction proceeds by refluxing the reactant and triethyl amine, a mild organic base, in an inert solvent such as tetrahydrofuran, acetonitrile.

The intermediate of Formula 5 is conveniently obtained by reacting a compound of Formula 7 with chloroacetyl chloride or bromoacetyl chloride.

[Formula 7]

In the above, R ₁ , R ₂ , R ₃ , R ₄ , R and A ₁ are as mentioned above. The reaction proceeds in a suitable solvent such as dichloromethane, 1,2-dichloroethane or tetrahydrofuran at room temperature in the presence of a gentle organic base such as triethyl amine.

Commercially available anthranilate esters of formula (7) and aromatic thiol compounds of formula (6) may be prepared by procedures described in the preparations described below, or by procedures similar to known compounds.

The manufacturing process of the compound of Formula 1 of the present invention can be summarized step by step as follows.

1) obtaining a compound of formula 5 by reacting a compound of formula 7 with chloroacetyl chloride or bromoacetyl chloride (first step),

2) reacting the compound of Formula 5 with the compound of Formula 6 to obtain a compound of Formula 4 (second step), and

3) cyclization of the compound of formula 4 to obtain a compound of formula 1 which is the target compound of the present invention (third step).

Hereinafter, the preparation method of the intermediate of the present invention will be described in detail by the preparation examples. Various conditions and parameters can be varied and applied in clinical treatment if it is clear that it is within the scope of the present invention.

Preparation Example 1 N- (3,5-dichlorophenyl) -2-hydroxyimino-acetamide

The mixture of water (50 mL), concentrated hydrochloric acid (12 mL) and 3,5-dichloroaniline (10.0 g, 61.7 mmol) in 1,4-dioxane (20 mL) was heated to a clear solution, which was then heated to 50 It was added to a mixture of water (224 mL), chloral hydrate (10.5 g, 66.9 mmol) and sodium sulfate (66.0 g) heated to < RTI ID = 0.0 > To the mixture was added hydroxylamine hydrochloride (13.0 g, 180 mmol) dissolved in 60 mL of water, and the mixture was refluxed for 50 minutes. After cooling to room temperature, the undissolved solid was filtered off, washed with excess water and dried under vacuum to give 12.8 g (89% yield) of the title compound as a pale yellow solid.

TLC R _f = 0.5 (EtOAc: n-hexane = 1: 3); mp 196-197 ° C .; ¹ H-NMR (DMSO-d ₆ ) δ 7.39 (t, J = 1.8 Hz, 1H, ArH), 7.70 (s, 1H, CHNOH), 7.89 (d, J = 1.8 Hz, 2H, ArH), 10.54 ( br s, 1H, NH), 12.40 (br s, 1H, NOH); MS (EI) m / e 233 [M ⁺ ], 216 [M ⁺ -OH], 202, 189, 161.

Preparation Example 2 4,6-dichloro-1H-indole-2,3-dione

To sulfuric acid (50 mL) in an ice bath was slowly added the compound (10.0 g) obtained in Preparation Example 1. At this time, the reaction mixture should be kept below 50 ° C. After complete addition, the black solution was heated to 90 ° C. for 10 minutes. After cooling the mixture to room temperature, the mixture was poured into crushed ice of 10 times the volume of the reaction solution and shaken vigorously for 1 hour. The undissolved solid formed was collected, washed with water and dried under vacuum to give 8.90 g (yield 96%) of the title compound as an orange solid.

TLC R _f = 0.4 (EtOAc: n-hexane = 1: 3); mp 228-230 ° C .; ¹ H-NMR (DMSO-d ₆ ) δ 6.97 (d, J = 1.8 Hz, 1H, ArH), 7.32 (d, J = 1.8 Hz, 1H, ArH), 11.42 (br s, 1H, NH); MS (EI) m / e 216 [M ⁺ ], 188 [M ⁺ -CO], 160.

Preparation Example 3 2-Amino-4, 6-dichlorobenzic acid

At room temperature, hydrogen peroxide (28% v / v, 10 mL) was added in portions to a solution of the compound (5.0 g, 23.1 mmol) obtained in Preparation Example 2 in 75 mL of 1N sodium hydroxide (aq). The mixture was stirred for 2 hours and filtered to remove undissolved dark brown solid. The yellow precipitate formed by carefully acidifying the filtrate with concentrated hydrochloric acid at pH 2, collected with water and dried under vacuum. Recrystallization with benzene gave 3.90 g (yield 82%) of the title compound as an ivory solid.

TLC R _f = 0.1 (EtOAc: n-hexane = 1: 1); mp 188-189 ° C; ¹ H-NMR (DMSO-d ₆ ) δ 6.76 (d, J = 1.9 Hz, 1H, ArH), 6.85 (d, J = 1.9 Hz, 1H, ArH); MS (EI) m / e 206 [M ⁺ ], 162 [M ⁺ -CO ₂ ].

Preparation Example 4 2-Amino-4.6-dichlorobenzic acid methyl ester

At ice bath temperature, a diazomethane ether solution was added portionwise to a solution of compound (11.5 g, 55.8 mmol) obtained in Preparation Example 3 in 20 mL of ethyl ether. After complete addition, the mixture was heated to room temperature and stirred until it dissolved completely. Thereafter, acetic acid (6.87 mL, 120 mmol) was added to the mixture to remove unreacted diazomethane, and then the solvent was evaporated under reduced pressure to give a yellow syrup, which was left to solid under vacuum to solidify orange. The title compound (12.1 g, 100% yield) was obtained as a solid.

¹ H-NMR (CDCl ₃ ) δ 3.88 (s, 3H, OCH ₃ ), 5.09 (br s, 2H, NH ₂ ), 6.56 (d, J = 1.8 Hz, 1H, ArH), 6.73 (d, J = 1.8 Hz, 1H, ArH); MS (EI) m / e 219 [M ⁺ −1], 190, 187.

Preparation Example 5 2-Amino-4-chlorobenzic acid methyl ester

In the same manner as in Preparation Example 4, the title compound was prepared using a commercially available 2-amino-4-chlorobenzic acid (20.0 g, 116.5 mmol) and a diazomethane ether solution. After usual treatment, the title compound (21.0 g, yield 97%) was obtained as an orange solid.

¹ H-NMR (CDCl ₃ ) δ 3.84 (s, 3H, CO ₂ CH ₃ ), 5.78 (br s, 2H, NH ₂ ), 6.57 (dd, J _A = 2.0 Hz, J _B = 8.6 Hz, 1H, ArH), 6.64 (d, J = 2.0 Hz, 1H, ArH), 7.75 (d, J = 8.6 Hz, 1H, ArH); MS (EI) m / e 184 [M ⁺ ], 126.

Preparation Example 6 3,5-dichloro-2- (2-chloro-acetylamino) -benzic acid methyl ester

Triethylamine (0.56 mL, 4.0 mmol) and chloroacetyl chloride (0.19 mL, 2.4 mmol) were added to the suspension of the compound (0.44 g, 2.0 mmol) obtained in Preparation Example 4 in anhydrous methylene chloride (CH ₂ Cl ₂ ). The mixture was stirred at room temperature for 4 hours in a nitrogen atmosphere. After the reaction was completed, the mixture was diluted with 100 mL of methylene chloride, washed with 1N hydrochloric acid and water, and dried over sodium sulfate. The solvent was then removed under reduced pressure to give a crude output. The crude product was recrystallized using methanol to give the title compound (0.55 g, yield 93%) as a white solid.

¹ H NMR (CDCl ₃ ) δ 3.99 (s, 3H, OCH ₃ ), 4.18 (s, 2H, COCH ₂ ), 7.25 (d, J = 2.0 Hz, 1H, ArH), 8.38 (d, J = 2.0 Hz , 1H, ArH); MS (EI) m / e 295 [M ⁺ ].

Preparation Example 7 4-Chloro-2- (2-chloro-acetylamino) benzic acid methyl ester

The title compound was prepared in the same manner as in Preparation Example 6, using the title compound (2.7 g, 14.6 mmol), chloroacetyl chloride (2.37 mL, 30.0 mmol) and triethylamine obtained in Preparation Example 5. After normal workup, recrystallization with methanol gave the pure title compound (3.44 g, yield 90%) as a white solid.

mp 134-135 ° C .; ¹ H-NMR (CDCl ₃ ) δ 3.96 (s, 3H, OCH ₃ ), 4.22 (s, 2H, CH ₂ ), 7.13 (dd, J = 2.1, 6.5 Hz, 1H, ArH), 7.99 (d, J = 6.5 Hz, 1H, ArH), 8.81 (d, J = 2.1 Hz, 1H, ArH), 11.92 (br s, 1H, NH); MS (EI) m / e 261 [M ⁺ ], 180.

Preparation Example 8 2- [2- (benzothiazol-2-ylsulfanyl) -acetyl amino] -4-chloro-benzic acid methyl ester

To a THF (15 mL) solution of the title compound (0.52 g, 2.00 mmol) obtained in Preparation Example 7 was added 2-mercapto benzothiazole (0.37 g, 2.20 mmol) and triethylamine (0.30 mL, 2.20 mmol). The mixture was refluxed for 5 hours and then washed with 5% NaHCO ₃ solution. And the organic layer was dried using MgSO ₄ and concentrated. This was recrystallized from ethyl acetate and hexane as a solvent to obtain the title compound (0.73 g, yield 90%) as a white solid.

mp 117-120 ° C .; ¹ H-NMR (CDCl ₃ ) δ 3.83 (s, 3H, OCH ₃ ), 4.26 (s, 2H, CH ₂ ), 6.99-7.92 (m, 6H, ArH), 8.77 (s, 1H, ArH), 11.72 (br s, 1 H NH); MS (EI) m / e 393 [M ⁺ ], 208, 180.

Preparation Example 9 2- [2- (benzoxazol-2-ylsulfanyl) acetyl amino] -4-chloro benzic acid methyl ester

Using the title compound (0.52 g, 2.00 mmol) and 2-mercaptobenzoxazole (0.31 g, 2.00 mmol) obtained in Preparation Example 7, the title compound (0.65 g, yield 86) was obtained in the same manner as in Preparation Example 8. %) Was obtained.

mp 120-122 ° C .; ¹ H-NMR (CDCl ₃ ) δ 3.81 (s, 3H, OCH ₃ ), 4.23 (s, 2H, CH ₂ ), 7.04-7.63 (m, 5H, ArH), 7.90 (d, J = 8.5Hz, 1H , ArH), 8.79 (d, J = 2.0 Hz, 1H, ArH), 11.75 (br s, 1H, NH); MS (EI) m / e 376 [M ⁺ ], 192, 165.

Preparation Example 10 4-Chloro-2- [2- (1-methyl-1H-imidazol-2-ylsulfanyl) acetyl amino] benzic acid methyl ester

The title compound (0.70) as a white solid, in the same manner as in Example 8, using the title compound (0.65 g, 2.50 mmol) and 2-mercapto-1-methylimidazole (0.28 g, 2.50 mmol) obtained in Preparation Example 7. g, yield 83%).

mp 93-95 ° C .; ¹ H-NMR (CDCl ₃ ) δ 3.67 (s, 3H, NCH ₃ ), 3.82 (s, 3H, OCH ₃ ), 4.01 (s, 2H, CH ₂ ), 6.91-7.09 (m, 3H, ArH), 7.93 (d, J = 8.5 Hz, 1H, ArH), 8.77 (d, J = 2.1 Hz, 1H, ArH), 11.57 (s, 1H, NH); MS (EI) m / e 339 [M ^< ⁺ &^gt;], 128, 95.

Preparation Example 11 2- [2- (1H-benzoimidazol-2-ylsulfanyl) acetyl amino] -4-chloro benzic acid methyl ester

Using the title compound (0.78 g, 3.00 mmol) and 2-mercaptobenzimidazole (0.50 g, 3.30 mmol) obtained in Preparation Example 7, the title compound (0.98 g, yield 87) was obtained in the same manner as in Preparation Example 8. %) Was obtained.

mp 146-147 ° C .; ¹ H-NMR (DMSO-d ₆ ) δ 3.86 (s, 3H, OCH ₃ ), 4.07 (s, 2H, CH ₂ ), 7.04-7.57 (m, 5H, ArH), 7.91 (d, J = 8.5 Hz , 1H, ArH), 8.75 (d, J = 2.1 Hz, 1H, ArH), 11.70 (s, 1H, NH); MS (EI) m / e 375 [M ⁺ ], 164.

Preparation Example 12 4-Chloro-2- [2- (1H- [1.2.4] triazol-3-ylsulfanyl) acetyl amino] -benzic acid methyl ester

Using the title compound (0.78 g, 3.00 mmol) and 1H-1.2.4-triazole-3-thiol (0.33 g, 3.26 mmol) obtained in Preparation Example 7, the title compound as a white solid was prepared in the same manner as in Preparation Example 8. 0.92 g, yield 94%) was obtained.

mp 174-175 ° C; ¹ H-NMR (DMSO-d ₆ ) δ 3.85 (s, 3H, OCH ₃ ), 4.07 (s, 2H, CH ₂ ), 7.24 (d, J = 6.5, 2.1 Hz, 1H, ArH), 7.94 (d , J = 8.5 Hz, 1H, ArH), 8.56 (br s, 2H, ArH), 11.35 (br s, 1H, NH); MS (EI) m / e 326 [M ⁺ +1], 185, 115.

Preparation Example 13 2- (3,5-Dichloro-2-methoxycarbonyl-phenylcarbamoylmethylsulfanyl) -nicotinic acid

Using the title compound (0.80 g, 2.69 mmol) and 2-mercaptonicotin acid (0.50 g, 2.82 mmol) obtained in Preparation Example 6, the title compound (1.05 g, yield 94%) was obtained in the same manner as in Preparation Example 8. )

mp 160 ° C .; ¹ H-NMR (DMSO-d ₆ ) δ 3.78 (s, 3H, CO ₂ CH ₃ ), 4.01 (s, 2H, COCH ₂ S), 7.28-7.34 (m, 1H, ArH), 7.57 (s, 1H , ArH), 7.77 (s, 1H, ArH), 8.28 (d, 1H, J = 7.8 Hz, 1H, ArH), 8.63 (d, J = 4.6 Hz, 1H, ArH), 10.11 (s, 1H, NH ).

Preparation Example 146- (3,5-dichloro-2-methoxycarbonyl-phenylcarbamoylmethylsulfanyl) nicotinic acid

The title compound (1.23) as a white solid, in the same manner as in Preparation Example 8, using the title compound (1.00 g, 3.37 mmol) and 6-mercaptonicotin acid (0.59 g, 90% tech., 3.54 mmol) obtained in Preparation Example 6. g, yield 88%).

mp 191-193 ° C; ¹ H-NMR (DMSO-d ₆ ) δ 3.78 (s, 3H, CO ₂ CH ₃ ), 4.19 (s, 2H, COCH ₂ S), 7.53-7.62 (m, 2H, ArH), 7.72 (d, J = 2.2 Hz, 1H, ArH), 8.13 (dd, J = 2.2, 8.2 Hz, 1H, ArH), 8.94 (d, J = 2.2 Hz, 1H, ArH), 10.29 (s, 1H, NH), 13.4 ( br s, 1H, COOH); MS (EI) m / e 415 [M ⁺ ], 384 [M ⁺ -OCH ₃ ], 366.

Preparation Example 15 2- [2- (5-Benzylcarbamoyl-pyridin-2-ylsulfanyl) -acetylamino] -4,6-dichloro-benzic acid methyl ester

Title compound (0.42 g, 1.01 mmol), 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide (0.72 g, 2.02 mmol) and 1-hydroxybentriazole (0.32 g, 2.02) obtained in Preparation Example 14. benzylamine (0.26 mL, 2.02 mmol) was added to a THF (30 mL) solution. The mixture was stirred for 2 days at room temperature, then diluted with EtOAc (50 mL), washed with 1N HCl 950 mL) and dried over MgSO ₄ . Thereafter, the solvent was evaporated and removed under vacuum in the mixture, and the mixture was separated by flash column chromatography (n-hexane: EtOAc = 9: 1) to obtain the title compound (0.31 g, 61% yield).

mp 270 ° C. decomp .; ¹ H-NMR (CDCl ₃ ) δ 3.78 (s, 3H, CO ₂ CH ₃ ), 3.98 (s, 2H, COCH ₂ S), 4.67 (d, J = 5.8 Hz, 2H, COCH ₂ NH), 6.43 ( br s, 1H, CONH), 7.18 (d, J = 2.0 Hz, 1H, ArH), 7.32-7.41 (m, 6H, ArH), 8.02 (dd, J = 2.0, 8.2 Hz, 1H, ArH), 8.26 (d, J = 2.2 Hz, 1H, ArH), 8.93 (d, J = 2.0 Hz, 1H, ArH), 10.07 (br s, 1H, NH).

Preparation Example 16 2- (5-Chloro-2-methoxycarbonyl-phenylcarbamoylmethylsulfanyl) -nicotinic acid

Using the title compound (0.84 g, 3.21 mmol) and 2-mercaptonicotin acid (0.51 g, 3.31 mmol) obtained in Preparation Example 7, the title compound (1.13 g, yield 93%) was obtained in the same manner as in Preparation Example 8. )

mp 165-167 ° C .; ¹ H-NMR (DMSO-d ₆ ) δ 3.86 (s, 3H, OCH ₃ ), 4.02 (s, 2H, CH ₂ ), 7.10-7.24 (m, 2H, ArH), 7.93 (d, J = 8.6 Hz , 1H, ArH), 8.23-8.67 (m, 3H, ArH), 11.44 (s, 1H, NH); MS (EI) m / e 196 [M ⁺ -185], 197, 185, 153.

Preparation 17 2,4-Dichloro-6- {2- [1- (4-hydroxy-phenyl) -1H-tetrazol-5-ylsulfanyl] -acetylamino} -benzic acid methyl ester

In the same manner as in Preparation Example 8, using the title compound (0.82 g, 2.75 mmol) and 1- (4-hydroxyphenyl) -1H-tetrazol-5-thiol (0.54 g, 2.78 mmol) obtained in Preparation Example 6. The title compound (1.15 g, yield 92%) was obtained as a brown solid.

mp 59-60 ° C. decomp .; ¹ H-NMR (CDCl ₃ ) δ 4.00 (s, 3H, OCH ₃ ), 4.16 (s, 2H, CH ₂ ), 6.99 (d, J = 8.7 Hz, 2H, ArH), 7.22 (d, J = 1.9 Hz, 1H, ArH), 7.41 (d, J = 8.8 Hz, 2H, ArH), 8.30 (d, J = 1.9 Hz, 1H, ArH).

Preparation 18 2,4-Dichloro-6- [2- (1H- [1.2.4] triazol-3-ylsulfanyl) -acetylamino] -benzic acid methyl ester

Using the title compound (0.60 g, 2.02 mmol) and 1H-1.2.4-triazole-3-thiol (0.22 g, 2.18 mmol) obtained in Preparation Example 6, the title compound as a white solid was prepared in the same manner as in Preparation Example 8. 0.68 g, yield 94%) was obtained.

mp 128-129 ° C .; ¹ H-NMR (CDCl ₃ ) δ 3.90 (s, 3H, OCH ₃ ), 3.92 (s, 2H, CH ₂ ), 7.21 (d, J = 1.8 Hz, 1H, ArH), 8.25 (br s, 2H, ArH); MS (EI) m / e 360 [M ⁺ ], 218, 115.

Hereinafter, the preparation method of the 3-heterocyclic mercapto-4-hydroxyquinolin-2 (1H) -one compound of the present invention will be described in detail by way of examples. However, the following examples are only for illustrating the present invention and the present invention is not limited by the examples.

Example 1 3- (benzothiazol-2-ylsulfanyl) -7-chloro-4-hydroxy-1H-quinolin-2-one

To a stirred solution of LiHMDS [prepared by treatment of hexamethyldisilazide (0.97 mL, 4.50 mmol) in n-BuLi (4.5 mmol) for 1 hour at −78 ° C. in freshly distilled THF (25 mL) at −78 ° C. A solution of the title compound of Preparation Example (0.52 g, 1.5 mmol) in freshly distilled THF (40 mL) was added dropwise. The reaction mixture was stirred for 1 hour and slowly warmed to room temperature. After further stirring for 4 hours, the reaction mixture was treated with trifluoroacetic acid until pH 1-2. The solvent was removed under reduced pressure to give a crude solid compound. The crude compound was washed with methanol to give the title compound (0.40 g, yield 75%) as a yellow solid.

mp 270 ° C .; ¹ H NMR (DMSO-d ₆ ) δ 7.27-7.49 (m, 4H, ArH), 7.81-8.02 (m, 3H, ArH), 11.90 (s, 1H, NH); MS (EI) m / e 360 [M ⁺ ], 179, 167.

Example 2 3- (benzoxazol-2-ylsulfanyl) -7-chloro-4-hydroxy-1H-quinolin-2-one

The title compound (0.60 g, 1.60 mmol) of Preparation Example 9 was obtained in the same manner as in Example 1, to obtain the title compound (0.23 g, yield 42%) as a brown solid.

mp 270 ° C .; ¹ H NMR (DMSOd ₆ ) δ 7.24-7.57 (m, 4H, ArH), 7.80-8.02 (m, 3H, ArH), 11.83 (br s, 1H, NH); MS (EI) m / e 344 [M ^< ⁺ &^gt;], 312, 163, 154.

Example 3 7-chloro-4-hydroxy-3- (1-methyl-1H-imidazol-2-ylsulfanyl) -1H-quinolin-2-one

The title compound (0.50 g, 1.47 mmol) of Preparation Example 10 was obtained in the same manner as the Example 1, to obtain the title compound (0.39 g, yield 86%) as a white solid.

mp 232-233 ° C; ¹ H NMR (DMSO-d ₆ ) δ 3.77 (s, 3H, NCH ₃ ), 7.09 (dd, J = 2.0, 6.6 Hz, 1H, ArH), 7.23 (d, J = 2.0 Hz, 1H, ArH), 7.43 (d, J = 1.9 Hz, 1H, ArH), 7.58 (d, J = 1.9 Hz, 1H, ArH), 7.87 (d, J = 8.5 Hz, 1H, ArH), 11.14 (br s, 1H, NH ); MS (EI) m / e 307 [M ⁺ ], 154, 126.

Example 4 3- (1H-benzoimidazol-2-ylsulfanyl) -7-chloro-4-hydroxy-1H-quinolin-2-one

The title compound (0.65 g, 1.73 mmol) of Preparation Example 11 was obtained in the same manner as in Example 1, to obtain the title compound (0.31 g, yield 52%) as a white solid.

mp 187-190 ° C .; ¹ H NMR (DMSO-d ₆ ) δ 7.12-7.41 (m, 6H, ArH), 7.90 (d, J = 8.6 Hz, 1H, ArH), 11.35 (br s, 1H, NH); MS (EI) m / e 343 [M ⁺ ], 313, 150.

Example 5 7-Chloro-4-hydroxy-3- (1H- [1.2.4] triazol-3-ylsulfanyl) -1H-quinolin-2-one

The title compound (0.45 g, 1.38 mmol) of Preparation Example 12 was obtained in the same manner as in Example 1, to obtain the title compound (0.35 g, yield 86%) as a white solid.

mp 270 ° C .; ¹ H NMR (DMSO-d ₆ ) d 7.24 (dd, J = 6.6, 2.0 Hz, 1H, ArH), 7.33 (d, J = 1.7 Hz, 1H, ArH), 7.90 (d, J = 8.7 Hz, 1H , ArH), 8.26 (s, 1H, ArH), 11.65 (s, 1H, NH); MS (EI) m / e 294 [M ⁺ ], 180.

Example 6 5,7-chloro-4-hydroxy-3- (nicotin-2-ylsulfanyl) -1H-quinolin-2-one

The title compound (0.42 g, 1.02 mmol) of Preparation Example 13 was obtained in the same manner as the Example 1, to obtain the title compound (0.23 g, 59% yield) as a white solid.

mp 270 ° C .; ¹ H NMR (DMSO-d ₆ ) δ 7.21-7.27 (m, 1H, ArH), 7.34-7.37 (m, 2H, ArH), 8.25 (d, J = 8.0 Hz, 1H, ArH), 8.43 (d, J = 4.8 Hz, 1H, ArH), 11.8 (s, 1H, NH).

Example 7 5,7-chloro-4-hydroxy-3- (nicotin-6-ylsulfanyl) -1H-quinolin-2-one

The title compound (0.30 g, 0.72 mmol) of Preparation Example 14 was obtained in the same manner as the Example 1, to obtain the title compound (0.23 g, yield 85%) as a white solid.

mp 275 ° C. decomp .; ¹ H NMR (DMSO-d ₆ ) δ 7.36 (d, J = 2.0 Hz, 1H, ArH), 7.40 (d, J = 2.2 Hz, 1H, ArH), 8.03 (dd, J = 2.4, 8.2 Hz, 1H , ArH), 8.84 (d, J = 1.6 Hz, 1H, ArH), 8.43 (d, J = 4.8 Hz, 1H, ArH), 11.94 (s, 1H, NH).

Example 8 5,7-chloro-4-hydroxy-3- (5-N-benzylcarbamoyl-pyridin-2-ylsulfanyl) -1H-quinolin-2-one

Using the title compound (0.31 g, 0.61 mmol) of Preparation Example 15 in the same manner as in Example 1, the title compound (0.17 g, 59% yield) was obtained.

mp 280 ° C. decomp .; ¹ H NMR (DMSO-d ₆ ) δ 4.45 (d, J = 5.6 Hz, 2H, ArCH ₂ NH), 7.13 (d, J = 8.2 Hz, 1H, ArH), 7.23-7.37 (m, 7H, ArH) , 8.01 (dd, J = 2.2, 8.4 Hz, 1H, ArH), 8.81 (d, J = 2.2 Hz, 1H, ArH), 9.12 (m, 1H, NH), 11.91 (s, 1H, NH).

Example 9 7-chloro-4-hydroxy-3- (nicotin-2-ylsulfanyl) -1H-quinolin-2-one

Using the title compound (0.22 g, 0.58 mmol) of Preparation Example 16 in the same manner as in Example 1, the title compound (0.08 g, yield 40%) was obtained.

mp 254-256 ° C. decomp .; ¹ H NMR (DMSO-d ₆ ) δ 7.18-7.33 (m, 3H, ArH), 7.87 (d, J = 8.7 Hz, 1H, ArH), 8.23 (d, J = 7.5 Hz, 1H, ArH), 8.40 (d, J = 4.6 Hz, 1H, ArH), 11.56 (s, 1H, NH).

Example 10 5,7-dichloro-4-hydroxy-3- [1- (4-hydroxy-phenyl) -1H-tetrazol-5-ylsulfanyl) -1H-quinolin-2-one

The title compound (0.63 g, 1.38 mmol) of Preparation Example 17 was obtained in the same manner as the Example 1, to obtain the title compound (0.52 g, yield 90%) as a white solid.

mp 250-252 ° C .; ¹ H NMR (DMSO-d ₆ ) δ 6.98 (d, J = 8.7 Hz, 2H, ArH), 7.33 (d, J = 1.8 Hz, 1H, ArH), 7.40 (d, J = 1.9 Hz, 1H, ArH ), 7.53 (d, J = 8.6 Hz, 2H, ArH), 10.28 (br s, 1H, OH), 11.94 (s, 1H, NH).

Example 11 5,7-dichloro-4-hydroxy-3- (1H- [1.2.4] triazol-3-ylsulfanyl) -1H-quinolin-2-one

The title compound (0.35 g, 0.97 mmol) of Preparation Example 18 was obtained in the same manner as the Example 1, to obtain the title compound (0.15 g, 47% yield) as a white solid.

mp 260-263 ° C. decomp .; ¹ H NMR (DMSOd ₆ ) δ 7.31 (s, 1H, ArH), 7.35 (s, 1H, ArH), 8.26 (br s, 1H, ArH), 11.8 (br s, 1H, NH); MS (EI) m / e 328 [M ^< ⁺ &^gt;], 296, 188.

I. In vitro screening for binding activity

1) Preparation of Synaptic Membrane

Male Sprague-Dawley rats (300-400 g) were obtained from a laboratory animal laboratory at Korea Research Institute of Chemical Technology. Laboratory animals were treated with water and standard in an air-conditioned room (22 + 1 ° C .; relative humidity, 60 + 5%) in a slightly darkened condition (brightness at 8 am) during the preparation period of 4-10 days prior to use. The laboratory food was fed and bred. Synaptic membranes for receptor binding studies are modified methods of Foster and Fagg [Eur. J. Pharmacol. 133, 291 (1987) and the modified method by Murphy et al. [Br. J. Pharmacol. 95, 932 (1988). In summary, male Sprague-Dawley rats were killed, brain cortex and hippocampus (chopocampus) were chopped with a scalpel and homogenized five times with 10 volumes of 0.32 M sucrose using a Teflon-fiber equalizer. Centrifuged for 10 minutes at 1000x g for 10 minutes with a Beckman J2 / 21 centrifuge (roto: JA20), and the supernatant was collected and centrifuged for 20 minutes at 20,000x g. The supernatant was removed and homogenized with a pellet homogenizer (5 times setting, 30 seconds).

After incubation at 4 ° C. for 30 minutes, the membrane suspension was centrifuged at 39,800 × g for 25 minutes with a Beckman L8-M ultracentrifuge. The pellet was left overnight at -70 ° C. The next day the pellet is dissolved at room temperature for 10 minutes and then resuspended in 20 volumes of 50 mM tris-acetone (pH 7.1, 4 ° C.) containing 0.04% Triton X-100, incubated at 37 ° C. for 20 minutes and 20 minutes. Centrifuge as above at 39,800 × g. The pellet was washed three times by centrifugation as above with 20 volumes of 50 mM tri-acetate at pH 7.1 without detergent. The final pellet was suspended in 50 mM Tris-Acetate, and protein concentration was measured using Bio-Rad agent (Bradford, 1976). The resuspension buffer was adjusted to a membrane protein concentration of 1 mg / ml and stored at -70 ° C.

2) [ ³ H] MDL-105,519 binding measurements

[ ³ H] MDL-105,519 binding was measured in 96-well plates. 50 μg of the synaptic membrane was added per well, and the final volume was 0.25 ml. The reaction mixture was incubated at 25 ° C. for 30 minutes with 50 mM tris-acetate buffer, pH 7.1. After varying concentrations of the test compound, a reaction mixture containing 4 nM of [ ³ H] MDL-105,519 was prepared and incubated as described above, and subjected to a drug displacement assay. After incubation, the reaction mixture is quickly filtered to terminate the reaction and inotech harvested using a Wallac GF / A glass fiber filter (Valax, Finland) pre-soaked in analytical buffer. ) Was washed nine times with ice-cold 50 mM Tris-acetone buffer 200. Radioactivity on the filter was measured by a plate counter (Wallac Microbeta) with a counting efficiency of 30-40%. Non-specific binding was measured in the presence of 1 mM glycine. All experimental compounds were dissolved in dimethylsulfoxide (DMSO) and diluted sequentially at various concentrations for binding measurements.

3) [ ³ H] MK-801 binding measurement

[ ³ H] MK-801 binding measurements are described by Wong et al., J. Neurochem. It was performed as described in 50, 274 (1988). For [ ³ H] MK-801 saturation binding assays, the synaptic membrane (300 μg of membrane protein) was prepared with 0.1-250 nM of [ ³ H] with or without 50 mM tris-acetate, 0.1 μM L-glutamate, pH 7.1. ] Incubated for 60 min at 30 ° C. in a final volume of 1 ml reaction mixture containing MK-801 and 1 μM glycine. For drug potential measurements, the synaptic membrane (membrane protein 200-300 μg) was placed in a reaction mixture containing 50 mM tris-acetate buffer at pH 7.1, 5 nM of [ ³ H] MDL-801 and various concentrations of experimental compound. It was incubated as above. The reaction was terminated as described above, filtered and counted for glycine binding assays. Nonspecific binding was measured in the presence of 0.1 mM (+) MK-801.

II. In vivo screening for anticonvulsive activity

1) NMDA (i.c.v.)-Induced spasm experiment

Based on the method of Chugai, 40-160 ng of NMDA per mouse was calculated using i.c.v. Obtained for induced spasms after administration. A 28 gauge injection needle attached to a 50 μl syringe was inserted through the bregma 1 mm right cartilage on the parietal suture and the test solution was inserted using a manipulator. The insertion volume was 5 μl per rat. Convulsions in response to NMDA occurred within 5 minutes, 1) rough running or jumping, 2) myoclonic seizures (isolated seizures and leg movements) and 3) (clonic seizures; loss of direction and repetitive movements of the limbs at the same time). Animals are i.c.v. Observation was performed for 10 minutes after injection and the seizure seen when myoclonus occurred. In order to investigate the NMDA-antagonist properties of the experimental drug, the following procedure was performed. The experimental drug (injection dose, 1 ml / 100 g) was i.p. 15 min after injection, 160 μl of NMDA was injected at 5 μl per horse. i.c.v. The experimental drug and NMDA solution were injected at the same time for investigation by administration. Ten rats were used for the same dose. The dose was increased until antagonistic NMDA reached a limit (eg solubility, lethality) that induces convulsions or can no longer be tested. NMDA was dissolved in 0.9% sodium chloride solution, and AP5,5,7-dichloroquinernic acid and 7-chloroquinernic acid were dissolved in a minimum amount of 0.1 N sodium hydroxide with 0.9% physiological saline.

2) Permanent Ligation of the Middle Celebral Atery (MCA)

Midbrain artery ligation is performed by Nagfuji et al. [Neurosci. Lett. 147, 159 (1992), Mol. Chem. Neuropathol. 26, 107 (1995), Neuro Report. 6, 1541 (1995). In summary, male Sprague Dawley rats were anesthetized with 2% halotan and then maintained at 1.5% halotan in nitrous oxide: oxygen (70:30) using a halotan evaporator. After the animal was lying on its side, the left temporal muscles were cut and partially separated from the iliac arc with bipolar coagulant. Small bilateral incisions were performed using a micro-drill cooled with cold saline in front of the hole of the mandible nerve under the surgical microscope. The medial end of the olfactory nerve and the middle cerebral artery were seen through the thin brain dura. The midbrain artery close to the lenticulostriate artery (LSA) was exposed and ligated using a miniclip. The midbrain artery and the lenticular striatum behind the miniclip were corroded with bipolar coagulant. During surgery, the temperature of the right temporal muscles and rectum was maintained at about 37 ° C. using a heating pad. The contracted temporal nerves were returned to place and sutured. The animals were placed in cages and kept warm using a heating lamp overnight. Twenty four hours after midbrain artery ligation, the animal's neck was cut quickly, soaked in cold saline, and sliced at 2 mm intervals from the frotal pole. The slices were freshly prepared in salt and incubated in 2% TTC solution preheated to 37 ° C. Slices chopped for 30 minutes at 37 ° C. were fixed with 4% formalin solution. An infarct region of each slice was measured with an image analyzer using a stereomicroscope. Infarct size in the left hemisphere was measured by subtracting the left hemisphere infarct size from the right hemisphere infarct size to rule out the effect of edema formation.

The 3-heterocyclic mercapto-4-hydroxyquinolin-2 (1H) -one derivative of the present invention is a potent and unique antagonist that acts on the trikinin non-sensitive glycine binding site in the NMDA receptor complex, It exhibits strong antagonistic activity against the central nervous system and has high solubility.

The compounds of the present invention are useful for the treatment or prevention of neurodegenerative diseases, and are particularly useful for reducing damage to the central nervous system caused by anemia or hypoxia such as heart attack, hypoglycemia, ischemia, cardiac arrest, trauma.

In addition, the compounds of the present invention are not only used for the prevention and treatment of chronic neurodegenerative diseases including epilepsy, Alzheimer's disease, Huntington's disease and Parkinson's disease, but also have an effect as antispasmodics, analgesics, antidepressants, anti-anxiety and antischizophrenia. have.

Claims (9)

3-heterocyclic mercapto-4-hydroxyquinolin-2 (1H) -one compound having the structure of formula 1, a tautomeric isomer thereof and a pharmaceutically acceptable salt thereof Formula 1 In the above, R represents a functional group represented by the following general formula (2) or (3). Formula 2 Formula 3 In Formula 2 or Formula 3 above, the closed circle represents two non-adjacent double bonds at any position of the pentagonal ring. A, B, C, and D independently represent oxygen, sulfur, nitrogen, or carbon, at least one of A, B, C, and D represents oxygen or sulfur, and at least one of A, B, C, and D represents carbon And others. E, F, G, H and I independently represent oxygen, sulfur, nitrogen or carbon, at least one of E, F, G, H and I represents oxygen or sulfur and E, F, G, H and I At least one of them represents something other than carbon. R ₁ , R ₂ , R ₃ and R ₄ are independently hydrogen, hydroxy, halogen, nitro, amino, haloalkyl, cyano, alkyl, alkenyl, alkynyl, azido, acylamino, aryl, alkoxy or hetero Represents a cycle ring. R ₅ and R ₆ are independently hydrogen, halogen, nitro, amino, carboxy, thiol, haloalkyl, cyano, alkyl, alkenyl, alkynyl, azido, acylamino, sulfonyl, aminosulfonyl, aryl, alkoxy , Heterocycle ring, acyloxy, alkylthio, arylthio, alkyl ester, alkyl carboxylate, aryl ester, aryl carboxylate, aralkyl ester, aralkyl carboxylate, urea or amidine. The compound of claim 1 wherein the compound is 3- (benzothiazol-2-ylsulfanyl) -7-chloro-4-hydroxy-1H-quinolin-2-one, 3- (benzoxazol-2-ylsulfanyl) -7-chloro-4-hydroxy-1H-quinolin-2-one, 7-chloro-4-hydroxy-3- (1-methyl-1H-imidazol-2-ylsulfanyl) -1H-quinolin-2-one, 3- (1H-benzoimidazol-2-ylsulfanyl) -7-chloro-4-hydroxy-1H-quinolin-2-one, 7-chloro-4-hydroxy-3- (1H- [1.2.4] triazol-3-ylsulfanyl) -1H-quinolin-2-one, 5,7-dichloro-4-hydroxy-3- (nicotin-2-ylsulfanyl) -1H-quinolin-2-one, 5,7-dichloro-4-hydroxy-3- (nicotin-6-ylsulfanyl) -1H-quinolin-2-one, 5,7-dichloro-4-hydroxy-3- (5-N-benzylcarbamoyl-pyridin-2-ylsulfanyl) -1H-quinolin-2-one, 7-chloro-4-hydroxy-3- (nicotin-2-ylsulfanyl) -1H-quinolin-2-one, 5,7-dichloro-4-hydroxy-3- (1H- [1.2.4] triazol-3-ylsulfanyl) -1H-quinolin-2-one 3-heterocyclic mercapto-4-hydroxyquinolin-2 (1H) -one compound, tautomer isomer thereof and a pharmaceutically acceptable salt thereof A pharmaceutical composition comprising the compound of claim 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. Use of the compound of claim 1, a pharmaceutically acceptable salt thereof, or a prodrug thereof as an antagonist of excitatory amino acids for the NMDA receptor. Use of the compound of claim 1, a pharmaceutically acceptable salt thereof, or a prodrug thereof to reduce damage to the central nervous system resulting from anemia or hypoxia such as stroke, hypoglycemia, ischemia, cardiac arrest, trauma. Use of the compound of claim 1, a pharmaceutically acceptable salt thereof, or a prodrug thereof as an agent for preventing and treating neurodegenerative diseases. Use of the compound of claim 1, a pharmaceutically acceptable salt thereof, or a prodrug thereof as a therapeutic agent for epilepsy, cerebral hemorrhage, Alzheimer's disease, Huntington's disease and Parkinson's disease. Use of the compound of claim 1, a pharmaceutically acceptable salt thereof, or a prodrug thereof as an anticonvulsant, analgesic, antidepressant, antianxiety and antischizophrenic agent. 1) obtaining a compound of formula 5 by reacting a compound of formula 7 with chloroacetyl chloride or bromoacetyl chloride (first step), 2) reacting the compound of Formula 5 with the compound of Formula 6 to obtain a compound of Formula 4 (second step), and 3) 3-heterocyclic mercapto-4-hydride of the present invention, comprising cyclization of a compound of Formula 4 to obtain a compound of Formula 1, which is the target compound of the present invention (Step 3). Method for preparing oxyquinolin-2 (1H) -one derivative. Formula 4 Formula 5 Formula 6 Formula 7